I am having difficulties showing glucose effects on human podocytes. In this sense, I am wondering if this is an effects of:
1) RPMI media contains 11mM glucose so "control" millieu is already too high;
Maybe people are buying RPMI with no glucose and then adding 5mM glucose? None of the papers I read said anything about it.
Yes, that is what I´m doing.
I´m buying RPMI-1640 with no glucose but I´m adding 5.5 mM glucose (low), and 25mM glucose as high glucose. I also have one group with Mannitol as control (5.5mM glucose medium + 19.5mM Mannitol).
Have you added ITS Liquid Media Supplement (I3146; Sigma Aldrich) in RPMI medium containing 11mM glucose during culturing and before the glucotoxicity experiment?
When I start my experiment, I change to only having insulin (10ug/mL).
I always have FBS and P/S in all media.
Good luck with your experiments.
Best,
Marie.
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